Journal ArticleDOI
Stimulation of lignocellulosic biomass hydrolysis by proteins of glycoside hydrolase family 61: structure and function of a large, enigmatic family
Paul Harris,Ditte Hededam Welner,Keith Mcfarland,Edward Re,Jens-Christian N. Poulsen,Kimberly Brown,Rune Salbo,Hanshu Ding,Elena Vlasenko,Sandy Merino,Feng Xu,Joel R. Cherry,Sine Larsen,Leila Lo Leggio +13 more
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TLDR
It is shown that certain GH61 proteins lack measurable hydrolytic activity but in the presence of various divalent metal ions can significantly reduce the total protein loading required to hydrolyze lignocellulosic biomass, and it is concluded that the GH 61 proteins are unlikely to be glycoside hydrolases.Abstract:
Currently, the relatively high cost of enzymes such as glycoside hydrolases that catalyze cellulose hydrolysis represents a barrier to commercialization of a biorefinery capable of producing renewable transportable fuels such as ethanol from abundant lignocellulosic biomass. Among the many families of glycoside hydrolases that catalyze cellulose and hemicellulose hydrolysis, few are more enigmatic than family 61 (GH61), originally classified based on measurement of very weak endo-1,4-beta-d-glucanase activity in one family member. Here we show that certain GH61 proteins lack measurable hydrolytic activity by themselves but in the presence of various divalent metal ions can significantly reduce the total protein loading required to hydrolyze lignocellulosic biomass. We also solved the structure of one highly active GH61 protein and find that it is devoid of conserved, closely juxtaposed acidic side chains that could serve as general proton donor and nucleophile/base in a canonical hydrolytic reaction, and we conclude that the GH61 proteins are unlikely to be glycoside hydrolases. Structure-based mutagenesis shows the importance of several conserved residues for GH61 function. By incorporating the gene for one GH61 protein into a commercial Trichoderma reesei strain producing high levels of cellulolytic enzymes, we are able to reduce by 2-fold the total protein loading (and hence the cost) required to hydrolyze lignocellulosic biomass.read more
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The paleozoic origin of enzymatic lignin decomposition reconstructed from 31 fungal genomes
Dimitrios Floudas,Manfred Binder,Robert Riley,Kerrie Barry,Robert A. Blanchette,Bernard Henrissat,Ángel T. Martínez,Robert Otillar,Joseph W. Spatafora,Jagjit S. Yadav,Andrea Aerts,Isabelle Benoit,Alexander Boyd,Alexis Carlson,Alex Copeland,Pedro M. Coutinho,Ronald P. de Vries,Patricia Ferreira,Keisha Findley,Brian Foster,Jill Gaskell,Dylan Glotzer,Paweł Górecki,Joseph Heitman,Cedar N. Hesse,Chiaki Hori,Kiyohiko Igarashi,Joel A. Jurgens,Nathan M Kallen,Phil Kersten,Annegret Kohler,Ursula Kües,T. K. Arun Kumar,Alan Kuo,Kurt LaButti,Luis F. Larrondo,Erika Lindquist,Albee Y. Ling,Vincent Lombard,Susan Lucas,Taina Lundell,Rachael Martin,David J. McLaughlin,Ingo Morgenstern,Emanuelle Morin,Claude Murat,László Nagy,Matthew J Nolan,Robin A. Ohm,Aleksandrina Patyshakuliyeva,Antonis Rokas,Francisco J. Ruiz-Dueñas,Grzegorz Sabat,Asaf Salamov,Masahiro Samejima,Jeremy Schmutz,Jason C. Slot,Franz J. St John,Jan Stenlid,Hui Sun,Sheng Sun,Khajamohiddin Syed,Adrian Tsang,Ad Wiebenga,Darcy Young,Antonio G. Pisabarro,Daniel C. Eastwood,Francis Martin,Dan Cullen,Igor V. Grigoriev,David S. Hibbett +70 more
TL;DR: Comparative analyses of 31 fungal genomes suggest that lignin-degrading peroxidases expanded in the lineage leading to the ancestor of the Agaricomycetes, which is reconstructed as a white rot species, and then contracted in parallel lineages leading to brown rot and mycorrhizal species.
Journal ArticleDOI
An oxidative enzyme boosting the enzymatic conversion of recalcitrant polysaccharides.
Gustav Vaaje-Kolstad,Bjørge Westereng,Svein Jarle Horn,Zhanliang Liu,Hong Zhai,Morten Sørlie,Vincent G. H. Eijsink +6 more
TL;DR: An enzyme is described that acts on the surface of crystalline chitin, where it introduces chain breaks and generates oxidized chain ends, thus promoting further degradation by chit inases, demonstrating the existence of a hitherto unknown enzyme activity.
Journal ArticleDOI
Expansion of the enzymatic repertoire of the CAZy database to integrate auxiliary redox enzymes
TL;DR: A new class introduced in the CAZy database is named “Auxiliary Activities” in order to accommodate a range of enzyme mechanisms and substrates related to lignocellulose conversion and provides a better coverage of the full extent of the lignin degradation machinery.
Journal ArticleDOI
Novel enzymes for the degradation of cellulose
TL;DR: This work describes recent developments in enzyme technology for conversion of cellulose, the most abundant, homogeneous and recalcitrant polysaccharide in lignocellulosic biomass, and focuses on a recently discovered new type of enzymes currently classified as CBM33 and GH61 that catalyze oxidative cleavage ofpolysaccharides.
Journal ArticleDOI
Insights into the oxidative degradation of cellulose by a copper metalloenzyme that exploits biomass components
R. Jason Quinlan,Matthew Sweeney,Leila Lo Leggio,Harm Otten,Jens-Christian N. Poulsen,Katja Salomon Johansen,Kristian B. R. M. Krogh,Christian Isak Jørgensen,Morten Tovborg,Annika Anthonsen,Theodora Tryfona,Clive P. Walter,Paul Dupree,Feng Xu,Gideon J. Davies,Paul H. Walton +15 more
TL;DR: It is demonstrated that copper is needed for GH61 maximal activity and that the formation of cellodextrin and oxidized cellodesxtrin products by GH61 is enhanced in the presence of small molecule redox-active cofactors such as ascorbate and gallate.
References
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